A clinical, genetic, and neuropathologic study in a family with 16q-linked ADCA type III.

Presented is the new kindred with autosomal dominant cerebellar ataxia linked to chromosome 16q22.1 (16q-ADCA type III) associated with progressive hearing loss. By haplotype analysis, the critical interval was slightly narrowed to three megabase regions between GATA01 and D16S3095. Neuropathologic study of 16q-ADCA type III demonstrated characteristic shrinkage of Purkinje cell bodies surrounded by synaptophysin-immunoreactive amorphous material containing calbindin- and ubiquitin-positive granules.

Application of vitrification to human embryo freezing.

To solve the problem of multiple pregnancies during the in vitro fertilization (IVF) and embryo transfer procedure, excess embryos must be cryopreserved for embryo transfer in future. We applied the vitrification method to cryopreservation of human embryos. A total of 31 frozen-thawed embryo transfer cycles were analyzed at the Yamagata University Hospital, Yamagata, Japan. The patients were introduced to IVF treatment and had an excess of valuable embryos to be frozen after the transfer of three fresh embryos that did not result in establishing a pregnancy. Excess human 8- to 16-cell stage embryos were exposed to vitrification solution and then frozen in liquid nitrogen. The cryoprotectant was removed by washing the embryos in media containing different concentrations of cryoprotectant. Three days after LH surge and/or 2 days after ultrasonographic ovulation the embryos were transferred. The rate of poor quality embryos significantly increased and the rate of good quality embryos decreased after thawing the embryos frozen by the vitrification method. In menstrual cycles with good quality embryo transfer, a higher rate of pregnancies was established than in the cycles in which fair or poor quality embryos were the highest grade of embryos transferred into the uterus. In total, 5 pregnancies were established from 31 embryo tansfers; 4 pregnancies were in cycles associated with the transfer of good quality embryos, and 1 pregnancy was in a cycle in which the highest grade of embryo was fair. When compared with slow embryo freezing methods, vitrification has marked advantages for clinical application in terms of cost and time. Vitrification will be an alternative method for embryo freezing.

Micturitional disturbances due to labial adhesion.

We report 2 cases of micturitional disturbances due to labial adhesion. Two women, aged 78 and 77 years old, visited our hospital with chief complaints of difficult voiding and urinary retention. On examination, their labias were found to be extensively fused. In both cases, the labial adhesions were separated by surgical repair under spinal anesthesia. Gentamicin and steroid ointments were used to prevent readhesion of the labia postoperatively.

Decreased membrane fluidity and unsaturated fatty acids in Niemann-Pick disease type C fibroblasts.

Niemann-Pick disease type C (NP-C) is an autosomal recessive disorder characterized by the sequestration and trapping of endocytosed cholesterol in lysosomes. The NPC1 gene on chromosome 18 was recently identified but its physiological function remains unknown. We have studied the lipid compositions of cultured human NP-C fibroblasts and mouse SPM-3T3 cell line derived from the C57BL/KsJ NP-C model mouse, which belongs to the same complementation group. Fibroblasts derived from apparently normal age-matched individuals and a subline of SPM-3T3 cells which restores cholesterol metabolism by transfer of human chromosome 18 were used as controls. Levels of free cholesterol in whole cell homogenates increased about 1.5-fold in human NP-C fibroblasts and mouse SPM-3T3 cells, while in the plasma membrane, cholesterol content did not significantly change in NP-C fibroblasts but rather decreased in SPM-3T3 cells. The total phospholipid content did not significantly change; however, among phospholipid head groups, increases in sphingomyelin and decreases in other classes were observed in human NP-C fibroblasts and mouse SPM-3T3 cells. The ratios of saturated fatty acids to unsaturated fatty acids increased in both human and mouse cells. The increase was also confirmed in the plasma membrane fraction of SPM-3T3 cells. Membrane fluidity was examined using a 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescent probe. The DPH anisotropy values were markedly increased in NP-C fibroblasts and in SPM-3T3 cells. The results suggest that a NP-C mutation causes complex alterations in cellular lipid contents and biophysical properties of the membrane.

Increased levels of GM2 ganglioside in fibroblasts from a patient with juvenile Niemann-Pick disease type C.

A 15-year-old boy was suffering from splenomegaly and a 10-year history of a neurologic disorder that included mental retardation, vertical supranuclear gaze palsy, dysarthria, ataxia, and dystonia. Bone marrow aspirates revealed foamy cells with storage materials which were positive with filipin staining. Cultured skin fibroblasts derived from the patient showed moderate loss of sphingomyelinase activity and the impairment of cholesterol esterification. The characteristic clinical presentations and typical histochemical findings of this patient met the diagnostic criteria of Niemann-Pick disease type C (NPC). In the fibroblasts from the patient, there was an accumulation of GM2 ganglioside around their cytoplasms. Increased levels of glycolipids. including GM2 ganglioside are reported in the cerebral cortex of NPC, but not in the fibroblasts. The fibroblasts derived from NPC may reflect the abnormal metabolism of glycolipids in the central nervous system of NPC.

The optimal equilibration time for mouse embryos frozen by vitrification with trehalose.

To determine the best equilibration time in the cryoprotectant before rapid cooling, 8-cell mouse embryos were exposed to a vitrification solution containing ethylene glycol, Ficoll and trehalose in modified phosphate-buffered saline at 5 degrees C for varying periods of time. They were frozen using an ultra rapid freezing method, thawed in a 20 degrees C water bath and cultured for 24 h with 5-bromo-2-deoxyuridine. Embryo development and the number of sister chromatid exchanges, a sensitive indicator of genetic damage, were observed. The results demonstrated that embryo development after freezing and thawing was similar among the groups exposed for periods of 5-40 min. However, the number of sister chromatid exchanges was significantly smaller in the group exposed for 5 min, indicating that this was the safest equilibration time in the vitrification solution.

Deafferentiation-induced c-fos gene expression in subthalamic nucleus and substantia nigra reticulata is reduced by non-NMDA receptor antagonist.

Molecular events underlying the mechanism by which brain injury elicits delayed transneuronal degeneration of neurons remote from the site of initial injury are not well understood. In rats, acute injury of the caudate nucleus (CN) and globus pallidus (GP) by local injection of excitotoxic ibotenic acid (IA) or by transient forebrain ischemia resulted in delayed cell death of neurons in the substantia nigra reticulata (SNr). To elucidate the involvement of glutamate receptor mediated hyperactivity of neurons produced by loss of inhibitory inputs in this delayed degeneration of SNr neurons, the region-specific expression of an immediate early gene, c-fos, and the effect of glutamate receptor antagonists on the c-fos expression were examined by using immunocytochemical and in situ hybridization analysis. Following unilateral IA-injection into the CN and GP, a robust expression of c-fos mRNA and Fos protein was induced specifically in neurons of both subthalamic nucleus (STN) and SNr deafferented by the IA-lesions 36 h after IA-injection. The delayed expression of Fos-protein in SNr neurons lasted for 48 h longer than that in STN neurons. Following unilateral IA-injection confined to the CN, an intense but short-term expression of Fos-protein was exhibited only in neurons of the deafferented SNr. c-fos mRNA and Fos protein were not expressed in neurons of the substantia nigra compacta at any time points examined. The induction of c-fos mRNA and Fos protein in neurons of the STN and SNr following IA-lesions of the CN and GP was reduced markedly by non-NMDA receptor antagonist (GYKI52466), but not by NMDA receptor antagonist (MK-801). The region-specific c-fos expression implies that deprivation of inhibitory afferents (disinhibition) due to destruction of presynaptic neurons can induce increased activity of postsynaptic neurons. The effect of GYKI52466 on the c-fos gene expression in neurons of the deafferented STN and SNr suggests that activation of non-NMDA receptors may be involved in a pathophysiological cascade for the transneuronal degeneration of SNr neurons.

Phase II study of intravesical chemoprophylaxis of epirubicin after transurethral resection of bladder tumors. Tottori University Oncology Group.

We performed intravesical instillation of epirubicin in 53 patients with the aim of preventing recurrence of bladder tumors. The patients had undergone transurethral resection of superficial bladder tumors (only transitional-cell carcinoma) within the preceding week, between January of 1990 and July of 1991. Recurrence was found in 11.3% (6/53) of cases during follow-up periods lasting from 1 to 20 months. Side effects occurred in 5.7% (3/53) of the patients. The cumulative nonrecurrence rate was 96.0% for a follow-up period of 6 months and 78.4% for 12 months. The results of this study suggest that intravesical instillation of epirubicin is very useful as adjuvant therapy after transurethral resection of superficial bladder tumors.

Effect of a centrally-acting muscle relaxant, eperisone hydrochloride, on muscle sympathetic nerve activity in humans.

The sympatho-modulating effects of eperisone hydrochloride, a centrally acting muscle relaxant, on microneurographically recorded muscle sympathetic nerve activity (MSA) were analyzed in human volunteers. A single dose of 300 mg of eperisone was orally administered to 19 healthy subjects aged between 19 and 27, and effects on 1) spontaneous MSA in 30 degrees head-up tilted position, 2) resting MSA and responsiveness to standing, 3) exercise-induced enhancement of MSA were observed. Eperisone has a sympatho-suppressive action in resting skeletal muscles, but has no effect on MSA in actively contracting muscles, e.g. standing, hand-gripping. The sympatho-suppressive effect of eperisone may be related to the drug-induced increase of blood low in the resting skeletal muscles; also it may be one more mechanism through which the drug can exert its muscle relaxant action.

Exteroceptive suppression of the trapezius muscle produced by mental nerve stimulation in normal subjects.

The purpose of the present study was to evaluate the similarities and differences in exteroceptive suppression produced by mental nerve stimulation between the masseter and trapezius muscles. Six normal subjects were studied using various intensities of stimulation. Although the duration and degree of exteroceptive suppression were increased with stimulus intensity in the masseter muscle, they did not correlate with stimulus intensity in the trapezius muscle. The latency and duration in the trapezius muscle were almost the same as those in the masseter muscle. The degrees in the masseter muscle were significantly larger than those in the trapezius muscle. Exteroceptive suppression of the trapezius muscle might have a similar mechanism as that of the masseter muscle, but it may be mediated by the separate interneurons. The projection from the trigeminal afferent to the trapezius motoneuron might be smaller than that to the masseter motoneuron.

Sympathetic nerve activity in the skin in relation to vibration-induced white finger.

With a microneurographic method, sympathetic nerve activity in the skin was recorded from median nerve fascicles of six patients with vibration-induced white finger and four age- and gender-matched normal subjects. At an ambient temperature of 15 degrees C the sympathetic nerve activity was greater in the patients than in the normal subjects. The burst activity was increased in all of the subjects during the water immersion test of the contralateral hand, but neither the increase in the mean burst number nor the mean area under the neurogram showed statistically significant group differences. Thus, although vasoconstrictive tone is greater in patients with vibration-induced white finger than in healthy subjects, no support could be found for the hypothesis that increased vascular tone is of prime importance for attacks of white finger in vibration-exposed subjects.

Long-acting alpha 1-adrenoceptive sympathomimetic agent suppresses sympathetic outflow to muscles in humans.

To clarify the effect of a long-acting selective alpha 1-adrenoceptive sympathomimetic agent on sympathetic outflow to muscles (muscle sympathetic activity, MSA) in humans, 5 mg of midodrine hydrochloride was injected intravenously in ten healthy male subjects aged 19-25. Spontaneous MSA was significantly suppressed as well as plasma norepinephrine level without changing heart rate, or systemic blood pressure. The suppression continued for more than 90 min, and this is supposed to be due to negative feedback action on the baroreflex loop. The absence of significant changes in heart rate or systemic blood pressure in healthy subjects suggests that hemodynamic homeostasis is maintained by reducing the MSA in response to a long-acting sympathomimetic agent, like midodrine hydrochloride, which constricts peripheral vessels directly.

Exteroceptive suppression of the masseter and temporalis muscles produced by electrical stimulation of the mental nerve in patients with Parkinson's disease.

Exteroceptive suppression of the masseter and temporalis muscles, produced by electrical stimulation of the mental nerve, was studied in 23 cases of Parkinson's disease (PD) and 20 age-matched control subjects. Two components were usually observed: early (ES1) and late (ES2). ES2 was estimated in this study. The background electromyographic activity prior to the stimulation showed no difference between normal subjects and patients with PD. The latency of suppression in patients with PD was not different from that in normal subjects. The duration and the degree of suppression were lower in patients with PD when compared with the normal subjects. The reduction of exteroceptive suppression might, at least in part, play a role in the failure of motor control of the face in PD.

[Exteroceptive suppression after mental nerve stimulation in patients with chronic headache].

In 18 normal subjects and 36 patients with chronic headache (3 cluster headache, 18 maigraine and 15 muscle contraction headache), exteroceptive suppression in the temporalis, masseter and trapezius muscles was examined. In the temporalis and masseter muscles, early and late phases (ES1 and ES2) were observed. In the present study, late response (ES2) was examined. Patients with muscle contraction headache showed an obvious decrease of exteroceptive suppression in duration and degree. Patients with common migraine showed slight reduction of exteroceptive suppression in duration and amount, when compared with normal subjects. Patients with classic migraine or cluster headache elicited almost normal exteroceptive suppression. Exteroceptive suppression in the trapezius muscle consisted of one or two suppressive phase(s). Early phase was small and unstable. In the present study, late response with latency of about 45 ms was examined. In the trapezius muscle, migraine and patients with muscle contraction headache elicited the reduction of the exteroceptive suppression in degree. There might be the failure of the inhibitory interneuron mediating the pathway of exteroceptive suppression not only in patients with muscle contraction headache, but also in patients with migraine, especially in patients with common migraine.

Skin nerve sympathetic activity reflex latency in Parkinson's disease.

Using a microneurographic method, skin nerve sympathetic activity (SSA) reflex latency was measured in 16 patients suffering from idiopathic Parkinson's disease (PD) and 11 age-matched normal subjects. The mean latency in patients with PD was significantly delayed (0.821 s, p less than 0.01), when compared with that in normal subjects (0.676 s). Reflex latency showed a significant positive linear correlation with arm length. By determining the SSA reflex latency, conduction velocities in post-ganglionic skin sympathetic nerve were measured indirectly. Averaged conduction velocities in PD and normal subjects, 1.76 m/s and 1.86 m/s, respectively, did not differ significantly (p greater than 0.1). This data suggests that the reflex pathway of SSA is disturbed in patients with PD and that the increased reflex latency is caused by a central/preganglionic delay.